For millennia, humankind has relied on plant derivatives for the prevention and treatment of a wide variety of aliments. For example, in China, various teas have been used as a crude medicine for over 4,000 years. And more recently, there has been considerable interest in taking advantage of various plant extracts as a source of health promoting substances such as, natural oxidants, flavonoids, and phenolic compounds. In part, this trend is due to a growing body of evidence demonstrating that some of these compounds have beneficial properties that may be advantageous in preventing or delaying, for example, the onset of cardiovascular disease.
Indeed, several studies have suggested that beneficial fatty acid and other plant derived compounds have desirable effects ranging from reducing lipid levels, lowering blood pressure, and regulating inflammatory disease. For example, barley has been shown to be particularly effective in lowering lipid levels in test animals (Quereshi et al., Lipids, 20:817-24 (1985)). And in particular, a tocochromanol isolated from barley extract has been identified as an active compound suitable for treating hypercholesterolemia (Quereshi et al., J. Biol. Chem., 261:10544-50 (1986)). Similarly, other tocochromanols, for example, xcex3-tocotrienol and xcex4-tocotrienol have also been shown to reduce hypercholesterolemia in mammals (European patent application 421,419).
In general, hypercholesterolemia involves high serum cholesterol levels that are associated with a number of diseases including atherosclerosis, arteriosclerosis, and cardiovascular disease. In addition, high serum cholesterol levels are also seen in patients suffering from other diseases such as diabetes mellitus and familial hypercholesterolemia. While improvement of lipoprotein profiles and a decrease in total serum and low density lipoprotein cholesterol have been shown to slow the progression of such diseases, the exact link between hypercholesterolemia and, most notably, cardiovascular disease, has remained obscure. As a result, cardiovascular disease continues to remain a leading cause of death in the United States.
In part, the reason a cure for cardiovascular disease has remained elusive, is that the etiology of the disease may be the result of series of complex interactions involving genetic factors, lipoprotein metabolism, clotting functions, and even lifestyle choices (e.g., diet, exercise). Interestingly, populations consuming large amounts of cereal grains have a lower incidence of cardiovascular disease and lower cholesterol levels. Studies looking at the beneficial properties of cereal diets have attributed these effects to naturally occurring tocochromanols, and these compounds have been found in a wide variety of plant sources (Quereshi et al., Am. J. Clin. Nutr., 53:1021S-6S (1991)).
As a class of compounds, tocochromanols include the tocopherols and the tocotrienols. Tocopherols, including xcex4- xcex1-tocopherol are essentially the active ingredient in vitamin E and have been extensively studied. A number of beneficial properties have been attributed to the tocopherols such as reduced platelet aggregation and antioxidant functions (Niki et al., Annals of the New York Academy of Sciences, 570:23-31 (1989); Fukuzawa et al., Annals of the New York Academy of Sciences, 570:449-453 (1989)).
The tocotrienols have been less well studied although recent evidence suggests that these compounds may also be biologically active (see for example U.S. Pat. No. 5,591,772 and 4,603,142). Naturally occurring tocotrienols including xcex1- xcex2-, xcex3-, and xcex4-tocotrienol have been identified in and isolated from a variety of sources including, e.g., rice, rice bran, barley, coconut, and palm. These compounds exhibit varying degrees of hypercholesterolemic activity and have also been used as antithrombotic agents and antioxidants.
Additional sources of tocopherols, tocotrienols, and other therapeutically beneficial compounds which can be used safely and effectively, for example, as a hypercholesterolemic, antithrombotic, antioxidizing, antiatherogenic, antiinflammatory, and immunoregulatory agents, would be of great benefit.
The present invention provides isolated cranberry seed oil, a novel source of health promoting compounds (e.g., desirable fatty acids, tocochromanols) that are useful in a variety of therapeutic applications, for example, as a hypocholesterolemic, antithrombotic, antioxidizing, antiatherogenic, antiinflammatory, and immunoregulatory agents. In addition, the invention provides methods of efficiently extracting cranberry seed oil to a high level of purity from cranberry seeds, for example, such that the extract can be added to foodstuffs or used as a dietary supplement or a pharmaceutical composition.
Accordingly, in one aspect, the invention provides an isolated cranberry seed oil extract which is substantially free of impurities. The extract can contain, for example, xcex1-tocopherol, xcex3-tocopherol, xcex4-tocopherol, xcex1-tocotrienol, xcex3-tocotrienol, xcex4-tocotrienol, or a combination thereof. In another embodiment, the extract further comprises an exogenous flavonoid, tamoxifen, or a combination thereof. In a related embodiment, the flavonoid is a flavone, flavavone, isoflavone, or flavonol. In another embodiment, the extract further comprises a fatty acid, preferably, xcex1-linolenic acid (omega-3), oleic acid (omega-9), linoleic acid (omega-6), or a combination thereof. In another embodiment, the extract further comprises a sterol, preferably, xcex2-sitosterol, schottenol (i.e., stigmastenol), or a combination thereof.
In another embodiment, the extract further comprises a triterpene alcohol, such as xcex1-amyrin, xcex2-amyrin, 24-methylene parkeol, or a combination thereof. In still another embodiment, the extract further includes a phenolic compound, preferably, methoxyphenylpropionic acid, methoxycinnamic acid, or a combination thereof.
In another aspect, the invention provides a therapeutic composition (e.g. a foodstuff, dietary supplement, or pharmaceutical composition) comprising isolated cranberry seed oil or one or a combination of the above listed compounds derived from cranberry seed oil. Accordingly, the composition can contain one or more of the following components: a tocochromanol (e.g., xcex1-tocopherol, xcex3-tocopherol, xcex4-tocopherol, xcex1-tocotrienol, xcex3-tocotrienol, xcex4-tocotrienol, or a combination thereof), an exogenous flavonoid (e.g., flavone, flavavone, isoflavone, or flavonol) a fatty acid (e.g., xcex1-linolenic acid (omega-3), oleic acid (omega-9), linoleic acid (omega-6), or a combination thereof), a sterol (e.g., xcex2sitosterol, schottenol (i.e., stigmastenol), or a combination thereof), a triterpene alcohol (e.g., xcex1-amyrin, xcex2-amyrin, 24-methyleneparkeol, or a combination thereof), or a phenolic compound (e.g., methoxyphenylpropionic acid, methoxycinnamic acid, or a combination thereof).
In another aspect, the invention provides a method for treating or preventing a disease or condition in a subject such as a malignancy, a hypercholesterolemic-related disease, a thrombotic disease, a respiratory disease, an atherogenic disease, an inflammatory disease or condition, a neurological disease, a dermatological disease, an opthalmological disease, or a gastroenterological disease, by administering to the subject a therapeutically-effective amount of a therapeutic composition (e.g., foodstuff, dietary supplement, or pharmaceutical composition) of the invention.
In a related embodiment, the subject has, or is at risk for acquiring, a malignancy, and is administered a composition comprising a tocotrienol, a flavonoid, tamoxifen, or a combination thereof.
In another related embodiment, the subject has or is at risk for acquiring a hypocholesterolemic-related disease, and is administered a composition comprising xcex1-tocopherol, xcex1-tocotrienol, xcex3-tocotrienol , xcex4-tocotrienol, or a combination thereof.
In yet another related embodiment, the subject has, or is at risk for acquiring, a respiratory disease, an inflammatory disease, a neurological disease, a derrnatological disease, an opthalmological disease, or a gastroenterological disease and is administered a composition comprising xcex1-tocopherol.
In another aspect, the invention provides a method for treating, preventing, or lowering the risk of acquiring a disorder or condition associated with an alteration in membrane stability, membrane fluidity, 5-lipoxygenase activity, or protein kinase C activity in a subject containing, the step of administering to the subject a therapeutically-effective amount of a therapeutic composition (e.g., foodstuffs, dietary supplements, or pharmaceutical compositions) of the invention.
In another aspect, the invention provides a method for nutritionally supplementing a foodstuff by adding to the foodstuff an isolated extract, or one or more components derived therefrom. Accordingly, in one embodiment, the foodstuff, comprises a tocochromanol (e.g., xcex1-tocopherol, xcex3-tocopherol, xcex4-tocopherol, xcex1-tocotrienol, xcex3-tocotrienol, xcex4-tocotrienol, or a combination thereof) an exogenous flavonoid (e.g., flavone, flavavone, isoflavone, or flavonol), tamoxifen, or a combination thereof, a fatty acid (e.g., xcex1-linolenic acid (omega-3), oleic acid (omega-9), linoleic acid (omega-6), or a combination thereof), a sterol (e.g., xcex2-sitosterol, schottenol (i.e., stigmastenol), or a combination thereof), a triterpene alcohol (e.g., xcex1-amyrin, xcex2-amyrin, 24-methylene parkeol, or a combination thereof), or a phenolic compound (e.g., methoxyphenylpropionic acid, methoxycinnamic acid, or a combination thereof).
In another aspect, the invention provides a method for isolating cranberry seed oil to a high level of purity, for example, so that the cranberry seed oil or components thereof can be administered to a patient or to an animal as a therapeutic. In one embodiment, the method involves physically disrupting cranberry seeds, adding to the seeds an organic solvent to produce an extract/solvent mixture, separating the extract/solvent mixture from the cranberry seeds, and removing the solvent portion of the extract/solvent mixture resulting in an isolated cranberry seed oil essentially free of solvent. In a preferred embodiment, the organic solvent is hexane and the adding step is conducted at a temperature between 50xc2x0 and 90xc2x0 F., and, more preferably, at a temperature between 50xc2x0 and 65xc2x0 F. In another embodiment, the separating step includes an extract receiver maintained at room temperature and atmospheric pressure. In another embodiment, the removing step is conducted at a temperature between 30xc2x0 and 220xc2x0 F. and under vacuum, preferably, a vacuum pressure of 22 inches of Hg or greater. In one embodiment, the extraction method results in an extraction yield by weight that is at least 10% or greater, preferably 15% or greater, and more preferably, at least 20% of the total weight of original seed or greater. In another embodiment, the extraction method also includes the step of increasing the oxidative stability of the resultant extract, by exposing the extract to ascorbic acid, BHT, low temperature, or a combination of these conditions.
In a related aspect, the invention provides isolated cranberry seed oil produced by the above-mentioned extraction method. Accordingly, the cranberry seed oil is substantially free of impurities. In one embodiment, the isolated cranberry seed oil contains a tocochromanol (e.g., a xcex1-tocopherol, xcex3-tocopherol, xcex2-tocopherol, xcex1-tocotrienol, xcex3-tocotrienol, xcex2-tocotrienol, or a combination thereof), an exogenous flavonoid, an exogenous tamoxifen, a fatty acid (e.g., xcex1-linolenic acid (omega-3), oleic acid (omega-9), linoleic acid (omega-6)), a sterol (e.g., xcex2-sitosterol, schottenol (i.e., stigmastenol), xcex1-amyrin, xcex2-amyrin, 24-methylene parkeol), a phenolic compound (e.g., methoxyphenylpropionic acid, methoxycinnamic acid, or a combination thereof) or any combination of the above components. In a related embodiment, the isolated cranberry seed oil is contained in a foodstuff, dietary supplement, or pharmaceutical composition. Accordingly, in another aspect, the invention provides a foodstuff, dietary supplement, or pharmaceutical composition comprising isolated cranberry seed oil, or one or more components thereof, produced by a method of the invention.